Anti-oxidative Stress Effect and Mechanism of Huangqintang on Caco-2 Cells Through Nrf2 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo verify the anti-oxidative stress effect of Huangqintang based on the nuclear factor E2-related factor 2 （Nrf2） signaling pathway by using Caco-2 cells as a carrier and RNA interference （RNAi） technology with in vitro experiments. MethodThe Caco-2 cells in the logarithmic growth phase were transfected with siRNA to construct siRNA Caco-2 cells. After normal Caco-2 cells and siRNA Caco-2 cells were incubated with Huangqintang of different doses， RNA and protein were extracted. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the mRNA and protein expression of heme oxygenase-1 （HO-1）， NAD（P）H quinone oxidoreductase 1 （NQO1）， glutathione S-transferase （GST）， Kelch-like ECH-associated protein 1 （Keap1）， and Nrf2. Meanwhile， the activities of superoxide dismutase （SOD） and GSH-Px， as well as the expression levels of malondialdehyde （MDA） and reactive oxygen species （ROS）， were detected by the colorimetric method and the probe method. ResultCompared with the results in the normal group， only the 400 mg·L-1 Huangqintang group and the sulforaphane （SFN） group could reduce the content of ROS and MDA in Caco-2 cells （P<0.01）， while the activities of SOD and GSH-Px in the cells of the Huangqintang groups and the SFN group showed an upward trend. Furthermore， there were significant differences in the 400 mg·L-1 Huangqintang group/the SFN group and the normal group （P<0.01）. Meanwhile， the protein and mRNA expression levels of HO-1， GST， Keap1， NQO1， and Nrf2 showed an upward trend in all groups （P<0.05， P<0.01）. After transfection， compared with the normal group， the model group showed increased content of MDA and ROS， blunted activities of GSH-Px and SOD， and reduced protein and mRNA expression of HO-1， GST， Keap1， and NQO1 （P<0.05， P<0.01）. After drug incubation， compared with the model group， the SFN group showed potentiated SOD activity， and the SFN group and the Huangqintang groups showed enhanced GSH-Px activity （P<0.01）. Moreover， the activities of SOD and GSH-Px in the 400 and 200 mg·L-1 Huangqintang groups and the SFN group showed an upward trend （P<0.01）， and the content of MDA in the 400 mg·L-1 Huangqintang group and the SFN group showed a downward trend. ROS decreased in all groups with drug intervention （P<0.01）， and the protein and mRNA expression of HO-1， GST， Keap1， NQO1， and Nrf2 increased to varying degrees （P<0.05， P<0.01）. ConclusionHuangqintang can play an anti-oxidative stress role by regulating the Nrf2 pathway.

Anti-inflammatory Effect of Huangqintang on LPS-induced RAW264.7 Inflammatory Cells / 中国实验方剂学杂志

ObjectiveTo explore the anti-inflammatory mechanism of Huangqintang based on the inflammation model in RAW264.7 cells. MethodHuangqintang was prepared and the safe dose to RAW264.7 cells was screened out. The RAW264.7 cells were seeded in 24-well plates and incubated with Huangqintang and lipopolysaccharide （LPS）， successively. The concentrations of nitric oxide （NO）， interleukin （IL）-6， tumor necrosis factor （TNF）-α， and prostaglandin E2 （PGE2） were measured by Griess assay and enzyme-linked immunosorbent assay （ELISA）， respectively. Meanwhile， RAW264.7 cells were inoculated in 6-well plates， and normal group， LPS group， LPS+Huangqintang group， nuclear factor-κB （NF-κB） p65 inhibitor PDTC group， p38 mitogen-activated protein kinase （MAPK） inhibitor SB203580 group， extracellular signal-regulated kinase （ERK） inhibitor PD98059 group， c-Jun N-terminal kinase （JNK） inhibitor SP600125 group， and Janus kinase （JAK） inhibitor AG490 group were set up. After the cells were incubated with corresponding inhibitors and Huangqintang and stimulated by LPS， RNA and protein were extracted. The mRNA and protein expression levels of NF-κB p65， p38 MAPK， ERK， JNK， and JAK were detected by Real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively， to explore the anti-inflammatory mechanism of Huangqintang by regulating the NF-κB， MAPK， and JAK/signal transducer and activator of transcription protein （STAT） signaling pathways. ResultAfter stimulation with LPS， the concentrations of NO， IL-6， TNF-α， and PGE2 in the cells of the model group increased significantly（P<0.05，P<0.01）. Compare with the model group， after incubation with Huangqintang， the secretion of NO， IL-6， TNF-α， and PGE2 showed a downward trend （P<0.05，P<0.01）. Compared with the normal group， the model group showed increased mRNA expression of p38 MAPK， ERK， JNK， JAK， and NF-κB p65 and total protein expression in cells after stimulation with LPS （P<0.05，P<0.01）. Compare with the model group，after incubation with Huangqintang， the total protein and mRNA expression of p38 MAPK， ERK， JNK， JAK， and NF-κB p65 in inflammatory cells decreased （P<0.05，P<0.01）. Meanwhile， the expression of NF-κB p65 total protein and mRNA in each inhibitor group showed a downward trend （P<0.05，P<0.01）. ConclusionHuangqintang can inhibit the inflammatory response through the NF-κB， MAPK， and JAK-STAT signaling pathways.

Effect of Huangqintang on Intestinal Flora in Mice with Ulcerative Colitis and Its Protective Mechanism on Intestinal Mucosal Barrier / 中国实验方剂学杂志

ObjectiveTo evaluate the pharmacodynamic effect of Huangqintang （HQT） on ulcerative colitis （UC） model mice and investigate its protective effect against UC by regulating intestinal flora. MethodMale Balb/c mice were randomly divided into control group，model group， high-， medium-， and low-dose HQT groups （20， 10， 5 g·kg-1）， flora interference group， flora interference model group， and flora interference-drug treatment group （HQT， 20 g·kg-1）. The flora interference model was constructed through intragastric administration of antibiotics （200 mg·kg-1 bacitracin and 200 mg·kg-1 vancomycin） for 8 d， and the UC model was constructed by allowing mice with free access to 3% dextran sulfate sodium （DSS） solution for 7 d. HQT was administered for 7 d. After the experiments， the mice were sacrificed， and blood， colon， and feces were collected. Hematoxylin-eosin （HE） staining was performed to observe the colonic lesions. The serum levels of interleukin （IL）-4， IL-6， IL-10， and tumor necrosis factor （TNF）-α were detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expression of Claudin1， MUC1， Occludin， and zonula occludens-1（ZO-1） in colon tissues was detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. The fecal DNA of mice was extracted and analyzed by high-throughput sequencing. ResultCompared with the normal group， the model group showed increased serum content of IL-4， IL-6， and TNF-α （P<0.05， P<0.01） and decreased IL-10 （P<0.05）. Compared with the model group， the HQT groups displayed decreased serum levels of IL-4， IL-6， and TNF-α （P<0.05， P<0.01）， increased IL-10 content （P<0.01）， increased mRNA and protein expression levels of Claudin1， MUC1， Occludin， and ZO-1 （P<0.05， P<0.01）. After flora interference， the diversity and abundance of intestinal bacteria decreased. To be specific， Proteobacteria increased （P<0.01）， and Firmicutes and Bacteroidetes decreased （P<0.01）. After UC induction by DSS， Bacteroidetes and Tenericutes decreased （P<0.05）. The high-， medium-， and low-dose HQT groups showed increased Bacteroidetes and Tenericutes （P<0.05， P<0.01） and decreased Firmicutes （P<0.05）. Additionally， the abundance of Lactobacillus， Lachnospiraceae NK4A136 group， Escherichia-Shigella， and Helicobacteris was positively proportional to the dose of HQT. ConclusionHQT can inhibit the inflammatory response of UC mice， restore the imbalance of intestinal flora， and repair the damaged intestinal mucosal barrier.

Mechanism of Huangqintang in Treatment of Ulcerative Colitis and Related Colon Cancer： A Review / 中国实验方剂学杂志

Ulcerative colitis （UC） is a chronic intestinal disease with unknown etiology， with main symptoms of abdominal pain， diarrhea， mucus， pus， and blood in the stool. It can be accompanied by various complications and has a high risk of developing to colon cancer. In recent years， the incidence of UC and related colon cancer has been increasing， which seriously affects human health and quality of life. The operation， immunosuppressant， etc. are the main approaches in the modern clinical treatment of UC and related colon cancer， but these methods all have different toxic and side effects， and the therapeutic effect is not ideal. For many years， traditional Chinese medicine （TCM） has attracted much attention in the treatment of UC and related colon cancer due to its slightly toxic side effects and remarkable curative efficacy. Huangqintang， derived from the Shang Han Lun （伤寒论）， is composed of Scutellariae Radix， Paeoniae Radix Alba， Glycyrrhizae Radix et Rhizoma， and Jujubae Fructus with the functions of clearing heat， checking diarrhea， harmonizing the middle， and relieving pain， and has a significant effect on the treatment of UC. Huangqintang has complex compositions and plays roles with multiple targets and pathways. According to the literature and the research results of this research group for many years， it was found that the mechanism of Huangqintang in the treatment of UC and related colon cancer was presumably related to the protection of the intestinal mucosal barrier， inhibition of inflammatory response， promotion of mitophagy， inhibition of oxidative stress， regulation of intestinal flora， cell cycle， and gene expression， suppression of cell proliferation， and promotion of apoptosis. To provide theoretical references for an in-depth study of the mechanism and clinical use of Huangqintang， this paper reviewed the research advances in recent years.

White Matter Damage in Major Depressive Disorder and TCM Treatment： A Review / 中国实验方剂学杂志

At present， major depressive disorder （MDD） is highly prevalent with advanced neurological disorders as the main pathological manifestations. As the physiological function bearer of higher neural activity， gray matter has become the focus of MDD treatment. However， recent research has shown that white matter and gray matter are independent of each other in the central nervous system （CNS）， and their functions are integrated and linked. In addition to gray matter damage， white matter damage is also the core driving event of disease progression and determines the outcome of MDD. At the treatment level， the current drug treatment of MDD mainly focuses on gray matter repair， while ignoring the importance of white matter integrity for the treatment of the disease， which has become the weakness of the current treatment of MDD. Traditional Chinese medicine （TCM） has good application potential in white matter repair. This paper elaborated on the following three aspects. ① The roles of white matter damage in the occurrence and development of MDD were summarized. ② The key link of white matter repair in MDD was elaborated with microglia microenvironment regulation as the entry point. ③ The application value of TCM in white matter repair in MDD was analyzed. This review aims to highlight the importance of white matter integrity in the treatment of MDD and is expected to expand the understanding dimension of the activity of related Chinese medicines in MDD from the perspective of white matter repair and analyze its potential application value.

Chinese perfusion practice survey results in 2021: current situation and challenge / 中华胸心血管外科杂志

Objective:To investigate the current situation of cardiopulmonary bypass(CPB) in China and analyze the causes, to guide the formulation and implementation of technology standard.Methods:The survey task force sent out a nationwide survey to obtain up-to-date information on perfusion practice by ChSECC(Chinese Society of Extracorporeal Circulation). The unit of analysis for the survey was the medical center performs CPB. The survey consisted 48 questions covering four topics of qualifications, including certification and education, policies and practices, device and equipment, techniques used.Results:There were 540 of the 714 centers for an overall response rate of 76%. According to the annual number of CPB, they were divided into 4 groups: group A(≤50 cases/year), group B(50-100 cases/year), group C(100-500 cases/year) and group D(≥500 cases/year). The response rate of center with more than group D last year was 100%. Most of the perfusionists had certification issued by ChSECC. Although there were more than 80% of group D performed regular training and assessment of perfusionist, the result was still not ideal enough. Low utilization of safety equipment was not depend on the annual operation volume in most of responding centers. Ultrafiltration and blood protection technology had high application rate in group D compared with group A and B.Conclusion:The certification rate of perfusionists are high. Lower the number of annual CPB cases, lower the proportion of regular evaluation and training, and lower rate of standards performance. No matter the amount of CPB, the application rate of safety equipment is not ideal. Higher the number of CPB cases, higher the utilization rate of CPB related technologies.

Evaluation of antibody responses to two-dose inactivated SARS-CoV-2 vaccines in patients with HIV-1 or chronic HBV infection: a single-center retrospective cohort study / 中华微生物学和免疫学杂志

Objective:To evaluate the efficacy and safety of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines in patients with HIV-1 or chronic HBV infection through observing the dynamic changes in antibody responses to two-dose inactivated SARS-CoV-2 vaccines.Methods:This cohort study recruited 169 people (including 39 with HIV-1 infection, 36 with chronic HBV infection and 94 individuals without chronic diseases) who completed two doses (prime and boost) of inactivated SARS-CoV-2 vaccination from January to December 2021. The levels of SARS-CoV-2 IgM and IgG antibodies at 14 d, one month and two months after boosting and neutralizing antibodies at one month were detected by chemiluminescence immunoassay and competitive ELISA method.Results:The positive rates of antibodies against SARS-CoV-2 in the HIV-1 and HBV groups were higher at one month after booster immunization, but significantly decreases at two months. The double-negative rate of SARS-CoV-2 IgM and IgG antibodies was higher in the HIV-1 and HBV groups than in the control group. The single positive rate of IgG antibody at one month in the control group was 2.01-fold higher than that of the HIV-1 group and 3.17-fold higher than that of the HBV group. The single positive rate of IgG antibody in people aged 18-39 years in each group was higher than that in the 40-59 age group. The antibody persistence was better in the HBV group than in the HIV-1 group, and the levels of IgG antibody in the HBV group was higher than that in the HIV-1 group. The neutralizing capacity of serum antibodies was significantly lower in the HIV-1 group than in the other groups ( P<0.000 1). The inhibition rate of serum neutralizing antibodies in the HBV group was lower than that in the control group among people aged 18-39 years [(34.050±6.031)% vs (64.220±3.845)%, t=4.43, P<0.000 1]. SARS-CoV-2-specific antibody responses were induced in 73.08% (19/26) of the patients aged 18-39 years in the HIV-1 group and 80.00% (4/5) in the HBV group. Conclusions:There were differences in the antibody responses to inactivated SARS-CoV-2 vaccines between different age groups, and infectious diseases affected the positive rates of antibodies and the neutralizing capability against SARS-CoV-2.

Risk factors of BK virus infection after kidney transplantation / 中华器官移植杂志

Objective:To retrospectively analyze the BKV infection of recipients after kidney transplantation(RT)and provide references for diagnosing and treating BK virus infection post-RT.Methods:From January 1, 2018 to December 31, 2020, clinical and follow-up data were reviewed for 561 RT recipients(cadaveric and living donor kidney)at First Hospital of Jilin University. DNA loading of BK virus in blood and urine was determined by quantitative polymerase chain reaction(qPCR)and kidney allograft biopsy performed. Based upon the results, they are divided into four groups of A (372 cases), high-level BK viruria(group B, 128 cases), BK viremia(group C, 52 cases)and BK virus nephropathy(BKVN)(group D, 9 cases). The variables related to BK virus infection were screened by univariate analysis. Meaningful variables( P<0.1)are incorporated into the multi-factor ordered Logistic regression model for examining the independent risk factors of postoperative BK virus infection. Results:The incidence of high-level BKV viruria is 33.69%(189/561)at 18 months post-RT. The average detection time is(4.2±3.8)months, the incidence of BK viremia 10.87%(61/561)and the average detection time(5.2±3.6)months post-RT. The incidence of BKVN is 1.78%(9/561)and the average detection time(7.0±4.0)months post-RT. Univariate analysis showed that gender, age, immunotherapeutic regimen, history of acute rejection and type of donor are correlated with BKV infection. Multivariate Logistic regression analysis indicated that male recipient( P=0.013), immune maintenance regimen( P<0.001)and history of acute rejection( P=0.002)were independent risk factors for developing postoperative BKV infection. Conclusions:There is a high incidence of BKV infection within 12 months post-RT. Male recipient, history of acute rejection and immune maintenance regimen are independent risk factors for BKV infection post-RT.

Efficacy and prognostic factors analysis of CT-guided 125I seeds implantation for primary hepatocellular carcinoma / 中华核医学与分子影像杂志

Objective:To evaluate the efficacy and prognostic factors of 125I seeds implantation for primary hepatocellular carcinoma. Methods:From December 2011 to January 2021, 102 primary hepatocellular carcinoma patients (86 males, 16 females; median age 61 years) who underwent 125I seeds implantation from 5 hospitals in China were enrolled in this retrospective study. Local progression-free survival (LPFS), overall survival (OS) and the prognostic factors were analyzed. Kaplan-Meier method was used to draw the distribution curve of survival time, and LPFS rate and OS rate were calculated. Log-rank test and Cox regression were used to analyze the influencing factors of survival. Results:The median follow-up time was 38 months until April 2021. The local control rate was 96.1%(98/102). The 1-, 3- and 5-year LPFS rate were 61.3%, 25.5% and 12.7%, and the 1-, 3- and 5-year OS rate were 73.9%, 39.1% and 22.6%, respectively. There were 75 patients with progressive disease, including 42 patients with intrahepatic recurrence and metastasis after seed implantation, and 55 patients died. Multivariate analyses showed that short-term efficacy complete response (CR) (hazard ratio ( HR)=0.34, 95% CI: 0.20-0.58) was protective factor related to LPFS; short-term efficacy CR ( HR=0.25, 95% CI: 0.13-0.47) was the protective factors related to OS; Barcelona clinic liver cancer (BCLC) C stage ( HR=2.33, 95% CI: 1.27-4.27), intrahepatic progression and extrahepatic metastasis ( HR=3.18, 95% CI: 1.28-7.86; HR=3.23, 95% CI: 1.27-8.21) were independent risk factors related to OS. No sever adverse effects were observed. Conclusions:125I seeds implantation is safe and effective for the treatment of primary hepatocellular carcinoma. BCLC stage, short-term efficacy and post-implantation progression are independent factors related to survival time.

Preparation and osteogenic properties of hydrogel scaffolds with different concentrations of laponite / 中华创伤骨科杂志

Objective:To prepare the hydrogel scaffolds with different concentrations of laponite and compare their osteogenic properties.Methods:The scaffolds of gelatin/sodium alginate hydrogel into which laponite was added according to the mass ratios of 0%, 1%, 2%, and 3% were assigned into groups T0, T1, T2, and T3. In each group, the compressive modulus was measured and the leaching solution for 24 h extracted to measure the ion release. Bone marrow mesenchymal stem cells (BMSCs) were cultured in the extract medium from each group and common medium (blank group) ( n=3) in the in vitro experiments to determine the expression of osteogenic genes Runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP) and type I collagen after 7 days of culture. In the in vivo experiments, the scaffolds were implanted into the femoral condyle defects in rats, and a blank group with no scaffolds was set. The bone repair in each group was evaluated by hematoxylin-eosin(HE) staining and immunohistochemical staining. Results:The compressive modulus in group T2 [(139.05±6.43) kPa] was significantly higher than that in groups T0, T1 and T3 [(68.83±3.76) kPa, (101.18±3.68) kPa and (125.40±3.28) kPa] ( P<0.05). The ion contents of lithium, magnesium and silicon released from the 24 h leaching solution in group T2 were (0.031±0.005) μg/mL, (3.047±0.551) μg/mL and (5.243±0.785) μg/mL, insignificantly different from those in group T3 ( P> 0.05) but significantly larger than those in group T1 ( P>0.05). The in vitro experiments showed that the expression levels of Runx2, ALP and type I collagen in group T2 were 1.59±0.11, 2.02±0.08 and 1.06±0.17, significantly higher than those in the other groups ( P<0.05). HE staining showed that the implanted hydrogel was tightly bound to the bone tissue. Immunohistochemical staining showed that the numbers of Runx2 and osteocalcin positive cells in group T2 were significantly higher than those in the other groups. Conclusions:With ideal biocompatibility, hydrogel scaffolds with different concentrations of laponite can slowly release the decomposed ions of lithium, magnesium and silicon to promote the osteogenic differentiation of BMSCs and the repair of bone defects in vivo. A 2% concentration of laponite in the hydrogel scaffolds may result in the best results.

Guideline for postoperative rehabilitation treatment following vertebral augmentation for osteoporotic vertebral compression fracture (version 2022) / 中华创伤杂志

Osteoporotic vertebral compression fracture (OVCF) can lead to lower back pain and may be even accompanied by scoliosis, neurological dysfunction and other complications, which will affect the daily activities and life quality of patients. Vertebral augmentation is an effective treatment method for OVCF, but it cannot correct unbalance of bone metabolism or improve the osteoporotic status, causing complications like lower back pain, limited spinal activities and vertebral refracture. The post-operative systematic and standardized rehabilitation treatments can improve curative effect and therapeutic efficacy of anti-osteoporosis, reduce risk of vertebral refracture, increase patient compliance and improve quality of life. Since there still lack relevant clinical treatment guidelines for postoperative rehabilitation treatments following vertebral augmentation for OVCF, the current treatments are varied with uneven therapeutic effect. In order to standardize the postoperative rehabilitation treatment, the Spine Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized relevant experts to refer to relevant literature and develop the "Guideline for postoperative rehabilitation treatment following vertebral augmentation for osteoporotic vertebral compression fracture (2022 version)" based on the clinical guidelines published by the American Academy of Orthopedic Surgeons (AAOS) as well as on the principles of scientificity, practicality and advancement. The guideline provided evidence-based recommendations on 10 important issues related to postoperative rehabilitation treatments of OVCF.

Advances in the study of structural modification and biological activities of asiatic acid / 药学学报

Asiatic acid (AA) is a ursane pentacyclic triterpenoids, which possesses a wide range of pharmacological activities, such as anti-tumor, hypoglycemic, anti-inflammatory, anti-bacterial. Due to poor solubility and low bioavailability, clinical application of asiatic acid is limited. To address these defects, the structural modifications of AA have been carried out, and large numbers of AA-based derivatives with novel structure and eximious biological activity have been developed. In this paper, the research progress of structural modifications, biological activity, structure-activity relationship and mechanism studies in recent twenty years are reviewed, which provides reference for development of AA-related drugs.

A nomogram for predicting lymph node metastasis in early gastric cancer / 中华胃肠外科杂志

Objective: To explore the independent risk factors of lymph node metastasis (LNM) in early gastric cancer, and to use nomogram to construct a prediction model for above LNM. Methods: A retrospective cohort study was conducted. Inclusion criteria: (1) primary early gastric cancer as stage pT1 confirmed by postoperative pathology; (2) complete clinicopathological data. Exclusion criteria: (1) patients with advanced gastric cancer, stump gastric cancer or history of gastrectomy; (2) early gastric cancer patients confirmed by pathology after neoadjuvant chemotherapy; (3) other types of gastric tumors, such as lymphoma, neuroendocrine tumor, stromal tumor, etc.; (4) primary tumors of other organs with gastric metastasis. According to the above criteria, 1633 patients with early gastric cancer who underwent radical gastrectomy at the Department of General Surgery of the Chinese PLA General Hospital First Medical Center from December 2005 to December 2020 were enrolled as training set, meanwhile 239 patients with early gastric cancer who underwent gastrectomy at the Department of General Surgery of the Chinese PLA General Hospital Fourth Medical Center from December 2015 to December 2020 were enrolled as external validation set. Risk factors of LNM in early gastric cancer were identified by using univariate and multivariate logistic regression analyses. A nomogram prediction model was established with significant factors screened by multivariate analysis. Area under the receiver operating characteristic curve (AUC) was used for assessing the predictive value of the model. Calibration curve was drawn for external validation. Results: Among 1633 patients in training set, the mean number of retrieved lymph nodes was 20 (13-28), and 209 patients (12.8%) had lymph node metastasis. Univariate analysis showed that gender, resection range, tumor location, tumor morphology, lymph node clearance, vascular invasion, lymphatic cancer thrombus, tumor length, tumor differentiation, microscopic presence of signet ring cells and depth of tumor invasion were associated with LNM (all P<0.05). Multivariate analysis revealed that females, tumor morphology as ulcer type, vascular invasion, lymphatic cancer thrombus, tumor length≥3 cm, deeper invasion of mucosa, and poor differentiation were independent risk factors for LNM in early gastric cancers (all P<0.05). Receiver operating characteristic curve indicated that AUC of training set was 0.818 (95%CI: 0.790-0.847) and AUC of external validation set was 0.765 (95%CI: 0.688-0.843). The calibration curve showed that the LNM probability predicted by nomogram was consistent with the actual situation (C-index: 0.818 in training set and 0.765 in external validation set). Conclusions: Females, tumor morphology as ulcer type, vascular invasion, lymphatic cancer thrombus, tumor length≥3 cm, deeper invasion of mucosa and poor differentiation are independent risk factors for LNM of early gastric cancer. The establishment of a nomogram prediction model for LNM in early gastric cancer has great diagnostic value and can provide reference for treatment selection.

Effects of non-muscle myosin Ⅱ silenced bone marrow-derived mesenchymal stem cells transplantation on lung extracellular matrix in rats after endotoxin/lipopolysaccharide-induced acute lung injury / 中华烧伤杂志

Objective: To investigate the effects of non-muscle myosin Ⅱ (NMⅡ) gene silenced bone marrow-derived mesenchymal stem cells (BMMSCs) on pulmonary extracellular matrix (ECM) and fibrosis in rats with acute lung injury (ALI) induced by endotoxin/lipopolysaccharide (LPS). Methods: The experimental research methods were adopted. Cells from femur and tibial bone marrow cavity of four one-week-old male Sprague-Dawley rats were identified as BMMSCs by flow cytometry, and the third passage of BMMSCs were used in the following experiments. The cells were divided into NMⅡ silenced group transfected with pHBLV-U6-ZsGreen-Puro plasmid containing small interference RNA sequence of NMⅡ gene, vector group transfected with empty plasmid, and blank control group without any treatment, and the protein expression of NMⅡ at 72 h after intervention was detected by Western blotting (n=3). The morphology of cells was observed by an inverted phase contrast microscope and cells labeled with chloromethylbenzoine (CM-DiⅠ) in vitro were observed by an inverted fluorescence microscope. Twenty 4-week-old male Sprague-Dawley rats were divided into blank control group, ALI alone group, ALI+BMMSC group, and ALI+NMⅡ silenced BMMSC group according to the random number table, with 5 rats in each group. Rats in blank control group were not treated, and rats in the other 3 groups were given LPS to induce ALI. Immediately after modeling, rats in ALI alone group were injected with 1 mL normal saline via tail vein, rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group were injected with 1×107/mL BMMSCs and NMⅡ gene silenced BMMSCs of 1 mL labelled with CM-DiⅠ via tail vein, and rats in blank control group were injected with 1 mL normal saline via tail vein at the same time point, respectively. At 24 h after intervention, the lung tissue was collected to observe intrapulmonary homing of the BMMSCs by an inverted fluorescence microscope. Lung tissue was collected at 24 h, in 1 week, and in 2 weeks after intervention to observe pulmonary inflammation by hematoxylin eosin staining and to observe pulmonary fibrosis by Masson staining, and the pulmonary fibrosis in 2 weeks after intervention was scored by modified Ashcroft score (n=5). The content of α-smooth muscle actin (α-SMA), matrix metalloproteinase 2 (MMP-2), and MMP-9 was detected by immunohistochemistry in 2 weeks after intervention (n=3), the activity of superoxide dismutase (SOD), malondialdehyde, myeloperoxidase (MPO) was detected by enzyme-linked immunosorbent assay at 24 h after intervention (n=3), and the protein expressions of CD11b and epidermal growth factor like module containing mucin like hormone receptor 1 (EMR1) in 1 week after intervention were detected by immunofluorescence staining (n=3). Data were statistically analyzed with one-way analysis of variance, Bonferroni method, and Kruskal-Wallis H test. Results: At 72 h after intervention, the NMⅡprotein expression of cells in NMⅡ silenced group was significantly lower than those in blank control group and vector group (with P values <0.01). BMMSCs were in long spindle shape and grew in cluster shaped like vortexes, which were labelled with CM-DiⅠ successfully in vitro. At 24 h after intervention, cell homing in lung of rats in ALI+NMⅡ silenced BMMSC group was more pronounced than that in ALI+BMMSC group, while no CM-DiⅠ-labelled BMMSCs were observed in lung of rats in blank control group and ALI alone group. There was no obvious inflammatory cell infiltration in lung tissue of rats in blank control group at all time points, while inflammatory cell infiltration in lung tissue of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group was significantly less than that in ALI alone group at 24 h after intervention, and alveolar wall turned to be thinner and a small amount of congestion in local lung tissue appeared in rats of the two groups in 1 week and 2 weeks after intervention. In 1 week and 2 weeks after intervention, collagen fiber deposition in lung tissue of rats in ALI alone group, ALI+BMMSC group, and ALI+NMⅡ silenced BMMSC group was significantly aggravated compared with that in blank control group, while collagen fiber deposition in lung tissue of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group was significantly improved compared with that in ALI alone group. In 2 weeks after intervention, modified Ashcroft scores for pulmonary fibrosis of rats in ALI alone group, ALI+BMMSC group, and ALI+NMⅡ silenced BMMSC group were 2.36±0.22, 1.62±0.16, 1.06±0.26, respectively, significantly higher than 0.30±0.21 in blank control group (P<0.01). Modified Ashcroft scores for pulmonary fibrosis of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group were significantly lower than that in ALI alone group (P<0.01), and modified Ashcroft score for pulmonary fibrosis of rats in ALI+NMⅡ silenced BMMSC group was significantly lower than that in ALI+BMMSC group (P<0.01). In 2 weeks after intervention, the content of α-SMA in lung tissue of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group were significantly decreased compared with that in ALI alone group (P<0.05 or P<0.01). The content of MMP-2 in lung tissue of rats in the 4 groups was similar (P>0.05). The content of MMP-9 in lung tissue of rats in ALI alone group was significantly increased compared with that in blank control group (P<0.01), and the content of MMP-9 in lung tissue of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group was significantly decreased compared with that in ALI alone group (P<0.01). At 24 h after intervention, the activity of malondialdehyde, SOD, and MPO in lung tissue of rats in ALI alone group, ALI+BMMSC group, and ALI+NMⅡ silenced BMMSC group were significantly increased compared with that in blank control group (P<0.01), the activity of malondialdehyde in lung tissue of rats in ALI+NMⅡ silenced BMMSC group and the activity of SOD in lung tissue of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group were significantly increased compared with that in ALI alone group (P<0.05 or P<0.01), and the activity of SOD in lung tissue of rats in ALI+NMⅡ silenced BMMSC group was significantly decreased compared with that in ALI+BMMSC group (P<0.01). The activity of MPO in lung tissue of rats in ALI+BMMSC group and ALI+NMⅡ silenced BMMSC group was significantly decreased compared with that in ALI alone group (P<0.01), and the activity of MPO in lung tissue of rats in ALI+NMⅡ silenced BMMSC group was significantly decreased compared with that in ALI+BMMSC group (P<0.01). In 1 week after intervention, the protein expression of CD11b in lung tissue of rats in ALI+NMⅡ silenced BMMSC group was significantly increased compared with those in the other three groups (P<0.05 or P<0.01), while the protein expressions of EMR1 in lung tissue of rats in the four groups were similar (P>0.05). Conclusions: Transplantation of NMⅡ gene silenced BMMSCs can significantly improve the activity of ECM components in the lung tissue in LPS-induced ALI rats, remodel its integrity, and enhance its antioxidant capacity, and alleviate lung injury and pulmonary fibrosis.

Analysis of influence factors for short-term recurrence of retroperitoneal liposarcoma after complete resection / 中华外科杂志

Objective: To examine the influence factors of short-term recurrence after complete surgical resection of retroperitoneal liposarcoma. Methods: The clinicopathological data of retroperitoneal liposarcoma at Department of General Surgery, the First Medical Center, People's Liberation Army General Hospital from January 2000 to January 2020 were retrospectively analyzed. There were 60 males and 31 females, aged (52.1±9.9) years (range: 30 to 84 years). Tumor recurrence within 12 months after complete resection was defined as short-term recurrence, and tumor recurrence more than 12 months was defined as non-short-term recurrence. The t test, rank-sum test, χ2 test and Fisher exact test were conducted for inter-group comparison. Logistic regression analysis was used to analyze the independent influence factors for the short-term recurrence of retroperitoneal liposarcoma after complete resection. The Kaplan-Meier curve was used to calculate the recurrence-free survival, and the Log-rank test was adopted for the comparison between the groups. Results: The univariate analysis results showed that irregular tumor morphology, multiple pathological subtypes, pathological scores>3, and multiple primary tumors are influence factors for short-term recurrence after complete resection of retroperitoneal liposarcoma (χ2: 4.422 to 7.773, all P<0.05). Regression analysis of the above risk factors showed that multiple primary tumors was the independent risk factor (OR=2.918, 95%CI: 1.127 to 7.556, P=0.027). In the short-term recurrence group, Kaplan-Meier curve analysis showed that patients with multiple primary tumors had a shorter median recurrence time than patients with unifocal tumor (6 months vs. 9 months, P=0.028). Conclusions: Multiple primary tumor is an independent risk factor for short-term recurrence after complete resection of retroperitoneal liposarcoma. It suggests that the frequency of follow-up after surgery should be increased for such patients.

Characteristics of fusion gene expression in acute lymphoblastic leukemia / 中华病理学杂志

Objective: To analyze the genetic landscape of 52 fusion genes in patients with de novo acute lymphoblastic leukemia (ALL) and to investigate the characteristics of other laboratory results. Methods: The fusion gene expression was retrospectively analyzed in the 1 994 patients with de novo ALL diagnosed from September 2016 to December 2020. In addition, their mutational, immunophenotypical and karyotypical profiles were investigated. Results: In the 1 994 patients with ALL, the median age was 12 years (from 15 days to 89 years). In the panel of targeted genes, 15 different types of fusion genes were detected in 884 patients (44.33%) and demonstrated a Power law distribution. The frequency of detectable fusion genes in B-cell ALL was significantly higher than that in T-cell ALL (48.48% vs 18.71%), and fusion genes were almost exclusively expressed in B-cell ALL or T-cell ALL. The number of fusion genes showed peaks at<1 year, 3-5 years and 35-44 years, respectively. More fusion genes were identified in children than in adults. MLL-FG was most frequently seen in infants and TEL-AML1 was most commonly seen in children, while BCR-ABL1 was dominant in adults. The majority of fusion gene mutations involved signaling pathway and the most frequent mutations were observed in NRAS and KRAS genes. The expression of early-stage B-cell antigens varied in B-cell ALL patients. The complex karyotypes were more common in BCR-ABL1 positive patients than others. Conclusion: The distribution of fusion genes in ALL patients differs by ages and cell lineages. It also corresponds to various gene mutations, immunophenotypes, and karyotypes.

Clinical study on 3% Diquafosol Sodium eye drops in treatment of meibomian gland dysfunction-related dry eye / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To observe the clinical effect of 3% diquafosol sodium eye drops in treatment of meibomian gland dysfunction-related dry eye.METHODS: The study involved 280 patients totally with meibomian gland dysfunction-related dry eye in the ophthalmology department, Hangzhou Hospital of Traditional Chinese Medicine from May 2020 to May 2021. Patients were divided into the treatment group(160 cases with 320 eyes)and the control group(120 cases with 240 eyes)according to the randomized number table method. The control group was treated with YangXueRunMu formula combined with 0.3% sodium hyaluronate eye drops, the treatment group was treated with YangXueRunMu formula combined with 3% diquafosol sodium eye drops. Both groups were administered for 4wk. The following indicators were measured before and after treatment at 2 and 4wk, respectively: the ocular surface disease index(OSDI)score, Schirmer I test( SⅠt), comprehensive analysis of tear meniscus height(TMH), non-invasive tear film break-up time(NITBUT), meibomian gland lipid secretion of smooth degree scoring and meibomian gland loss rate score, the determination of interleukin-6(IL-6)in tears and the level of tumor necrosis factor-alpha(TNF-α). The efficacy of these tests results was evaluated among these indicators.RESULTS: The overall effective rates of the treatment group and the control group were 95.6% and 81.7% respectively(P<0.05). After 2, 4wk of treatment, the ocular surface disease index(OSDI), NITBUT, meibomian gland lipid secretion scoring, meibomian gland loss rate score and the levels of IL-6 and TNF-α in tears of two groups were significantly different than before treatment(P<0.05). and the treatment group was better than the control group; there was no difference between the SⅠt and TMH groups before and after treatment in the two groups(P>0.05).CONCLUSION: The 3% diquafosol sodium eye drops can promote the normal secretion of meibomian gland by prolonging the homeostasis of the tear membrane, and it can also inhibit the release of inflammatory factors in tears in the treatment of blebomian gland dysfunction-related dry eye.

Study on key amino acids in E2 protein of recombinant side chain of dioxygate dehydrogenase complex in primary biliary cholangitis / 中华风湿病学杂志

Objective:To explore the key amino acids of side chain dioxate dehydrogenase complex E2 protein (BCOADC-E2) that can react specifically with specific autoantibody anti-mitochondrial antibodies (AMA)-M2 in patients with primary biliary cholangitis (PBC).Methods:The homologous target gene BCKD, which expressed the epitopes of BCOADC-E2 protein, was cloned and recombined with the engineering plasmid pGEX-4T1. Fifteen point mutant plasmids were obtained by polymerase chain reaction (PCR) and transferred to the prokaryotic expression strain for protein expression and purification. Fourteen mutant proteins and one wild-type protein were obtained. The AMA-M2 specificity of the 14 mutant proteins was measured by enzyme-linked immuno sorbent assay (ELISA), and the amino acids that were critical to the specificity of BCOADC-E2 and AMA-M2 were identified by comparing the specificity of the 14 mutant proteins with that of the wild type proteins. Differences between groups were analyzed by analysis of variance, LSD- t test. Results:A total of 14 mutant proteins and 1 wild-type protein were obtained.The specific reaction degree of mutant protein pGEX-BCKD-S1A (1.634±0.328) and pGEX-BCKD-C3A (1.744±0.345) with serum AMA-M2 in patients with PBC was higher than that of wild-type protein pGEX-BCKD (1.000±0.000) with AMA-M2; Mutant protein pGEX-BCKD-E4A (0.157±0.067), pGEX-BCKD-V5A (0.057±0.029), pGEX-BCKD-Q6A (0.580±0.166), pGEX-BCKD-S7A (0.744 ±0.125), pGEX-BCKD-D8A (0.351 ±0.135), pGEX-BCKD-S10A (0.496 ±0.158), pGEX-BCKD-V11A(0.149±0.089), pGEX-BCKD-T12A(0.061±0.043), pGEX-BCKD-I13A(0.007±0.017), pGEX-BCKD-T14A (0.198±0.101), pGEX-BCKD-S15A (0.156±0.087), The specific reaction degree of pGEX-BCKD-R16A (0.884±0.099) with AMA-M2 was lower than that of wild-type protein pGEX-BCKD (1.000±0.000) with AMA-M2.Conclusion:The cysteines at positions 1 and 3 of BCOADC-E2 protein were the key amino acids to improve the specific reaction between BCOADC-E2 and AMA-M2. The mutant proteins formed after amino acids at position 4, 5, 6, 7, 8, 10, 11, 12, 13, 14, 15 and 16 are replaced by alanine can decrease the specificity of AMA-M2. Amino acids at positions 5 and 13 are the key amino acids that affect the specific reaction between BCOADC-E2 and AMA-M2, and have an important effect on the function of BCOADC-E2 protein.

Xuebijing Injection Regulates Mitochondrial N-formyl Peptides/NLRP3 Inflammatory Pathway to Treat Severe Acute Pancreatitis in Rats / 中国实验方剂学杂志

ObjectiveTo investigate the therapeutic effect of Xuebijing injection （XBJ） on sodium taurocholate （Na-Tc）-induced severe acute pancreatitis （SAP） in rats. MethodForty rats were randomly assigned into 5 groups： sham operation group， SAP model group， and low-， medium-， and high-dose （4， 8， 12 mL·kg·d-1， respectively） XBJ groups. SAP model was established by retrograde injection of Na-Tc （1 mL·kg-1） into the biliary and pancreatic ducts. XBJ was injected intraperitoneally 3 days before and 0.5 h after modeling. The ascitic fluid volume and the pancreas weight-to-body weight ratio were measured. The pathological changes of pancreatic tissue were observed via hematoxylin-eosin （HE） staining. The protein levels of formyl peptide receptor 1 （FPR1） and nucleotide-binding oligomerization domain-like receptor 3 （NLRP3） in pancreatic tissue were detected by immunohistochemistry. Western blot was employed to determine the expression levels of NADH-ubiquinone oxidoreductase chains 1-6 （MT-ND1， MT-ND2， MT-ND3， MT-ND4， MT-ND5， and MT-ND6） in rat plasma. ResultCompared with sham operation group， the SAP model group showcased increased ascitic fluid volume and pancreas weight-to-body weight ratio （P<0.05）， serious lesions in pancreatic tissue， increased total pathological score （P<0.05）， and up-regulated protein levels of FPR1 and NLRP3 in pancreatic tissue （P<0.05）. The model group had lower MT-ND2 level （P<0.05） and higher MT-ND1， MT-ND3， and MT-ND6 levels in plasma （P<0.05） than the sham operation group， while MT-ND4 and MT-ND5 had no significant differences between the two groups. Compared with SAP model group， the XBJ treatment decreased ascitic fluid volume and pancreas weight-to-body weight ratio （P<0.01）， ameliorated pancreatic lesions， and down-regulated the protein levels of FPR1 and NLRP3 in pancreatic tissue （P<0.01）. The treatments， especially high-dose XBJ （P<0.01）， down-regulated the expression of MT-ND1 （P<0.01）， MT-ND3 （P<0.01）， MT-ND6 （P<0.01）， and MT-ND4 and did not change that of MT-ND5. ConclusionXBJ may antagonize partial mitochondrial N-formyl peptides and excessive inflammatory response mediated by FPR1/NLRP3 to treat SAP in rats.

Total Ginsenoside Extract from Panax ginseng Enhances Neural Stem Cell Proliferation and Neuronal Differentiation by Inactivating GSK-3β / 中国结合医学杂志

OBJECTIVE@#To study the effects of total ginsenosides (TG) extract from Panax ginseng on neural stem cell (NSC) proliferation and differentiation and their underlying mechanisms.@*METHODS@#The migration of NSCs after treatment with various concentrations of TG extract (50, 100, or 200 µ g/mL) were monitored. The proliferation of NSCs was examined by a combination of cell counting kit-8 and neurosphere assays. NSC differentiation mediated by TG extract was evaluated by Western blotting and immunofluorescence staining to monitor the expression of nestin and microtubule associated protein 2 (MAP2). The GSK-3β/β-catenin pathway in TG-treated NSCs was examined by Western blot assay. The NSCs with constitutively active GSK-3β mutant were made by adenovirus-mediated gene transfection, then the proliferation and differentiation of NSCs mediated by TG were further verified.@*RESULTS@#TG treatment significantly enhanced NSC migration (P<0.01 or P<0.05) and increased the proliferation of NSCs (P<0.01 or P<0.05). TG mediation also significantly upregulated MAP2 expression but downregulated nestin expression (P<0.01 or P<0.05). TG extract also significantly induced GSK-3β phosphorylation at Ser9, leading to GSK-3β inactivation and, consequently, the activation of the GSK-3β/β-catenin pathway (P<0.01 or P<0.05). In addition, constitutive activation of GSK-3β in NSCs by the transfection of GSK-3β S9A mutant was found to significantly suppress TG-mediated NSC proliferation and differentiation (P<0.01 or P<0.05).@*CONCLUSION@#TG promoted NSC proliferation and neuronal differentiation by inactivating GSK-3β.